Quality control of astrocyte-directed Cre transgenic mice: the benefits of a direct link between loss of gene expression and reporter activation.
Cre recombinase activity for cell-type restricted deletion of floxed target genes (i.e., flanked by Cre recognition loxP-sites) is often measured by separate matings with recombination-activated reporter gene mice. Using a floxed Gja1 (Cx43) allele, we demonstrate the benefits of a direct link between reporter gene expression and target gene deletion to overcome critical limitations of the Cre/loxP system. The widely used human glial fibrillary acidic protein (hGFAP)-Cre transgene exhibits variable recombination activity and requires postexperimental validation. Such quality control is essential to correlate the extent of Cre-mediated Gja1 ablation with phenotypical alterations and to maintain the activity status of hGFAP-Cre in transgenic mouse colonies. We present several strategies to control for the fidelity of hGFAP-Cre mediated recombination. (c) 2008 Wiley-Liss, Inc.
Requardt
Robert Pascal
RP
Institute of Genetics, Division of Molecular Genetics, University of Bonn, D-53117 Bonn, Germany.
Kaczmarczyk
Lech
L
Dublin
Pavel
P
Wallraff-Beck
Anke
A
Mikeska
Thomas
T
Degen
Joachim
J
Waha
Andreas
A
Steinhäuser
Christian
C
Willecke
Klaus
K
Theis
Martin
M
D000818:Animals; D001253:Astrocytes / Q000502:physiology; D017630:Connexins / Q000235:genetics; D005260:Female; D017353:Gene Deletion; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D008297:Male; D008745:Methylation; D051379:Mice; D008822:Mice, Transgenic; D011401:Promoter Regions, Genetic / Q000502:physiology; D011786:Quality Control
D017630:Connexins; D005904:Glial Fibrillary Acidic Protein